摘要:

We have developed a new resin for peptide synthesis that can be used to synthesize and evaluate directly combinatorial peptide libraries for binding target proteins. Fidelity of the peptide synthesis using this hydrophilic resin is comparable to polystyrene‐based resins. Peptide libraries synthesized on this resin were probed by a two color PEptide Library Immunostaining Chromatographic ANalysis (PELICAN) technique for sequences binding the serine protease Factor IX zynogen. This PELICAN technique readily distinguishes between beads interacting with the reagents for target detection (blue beads) from those beads specific for the target protein itself (red beads). Validation of the PELICAN technique, as well as purification of Factor IX from plasma, is demonstrated utilizing this resin. ©Munksgaard 19

ISSN:0367-8377    

DOI:10.1111/j.1399-3011.1996.tb00812.x

出版商:Blackwell Publishing Ltd    

年代:1996

数据来源: WILEY

摘要:

The interaction of the κ‐opioid receptor‐selective heptadecapeptide dynorphin A(1‐17) (Tyr1‐Gly‐Gly‐Phe‐Leu5‐Arg‐Arg‐Ile‐Arg‐Pro10‐Lys‐Leu‐Lys‐Trp‐Asp15‐Asn‐Glu) with phospholipid membranes has been investigated by monitoring the leakage of the internal aqueous contents of liposomes, the changes in the tryptophan emission spectrum, and the collisional quenching of tryptophan fluorescence by brominated lipids. The peptide induces more extensive leakage of contents from phosphatidylserine than from phosphatidyl‐choline vesicles, and experiences a blue shift of the Trp fluorescence emission maximum in the presence of phosphatidylserine vesicles. In the presence of phosphatidylcholine vesicles, however, the Trp fluorescence intensity is reduced without a blue shift. In phosphatidylserine membranes containing 10 mol% phosphatidylcholine, the intensity of the blue‐shifted fluorescence is enhanced. This avid interaction of dynorphin A(1‐17) with phosphatidylserine membranes is likely to be mediated by the positively charged Arg and Lys groups. It is proposed that. while theN‐terminus of the peptide may be embedded in the bilayer in analogy with dynorphin (1–13), theC‐terminal region of dynorphin A (1‐17) bends back onto the bilayer/water interphase, and that the Trp14, residue is stabilized in a hydrophobic pocked near the interphase by the interaction of the neighboring charged amino acids with the phosphate, carboxyl and

ISSN:0367-8377    

DOI:10.1111/j.1399-3011.1996.tb00813.x

出版商:Blackwell Publishing Ltd    

年代:1996

数据来源: WILEY

摘要:

Production of multiple overlapping peptides is a key step in the identification of T‐cell epitopes. A large number of peptides can be produced by using ABIMED's automated multiple peptide synthesizer. We report here considerable improvement in the software and chemistry of peptide synthesis by introducing a resin mixing step during coupling, when using this synthesizer. A comparison of two solvent systems for synthesis was performed. Six test peptides were synthesized by standard and modified methods. The purity of peptides, assessed by HPLC and mass spectrometry, showed a substantial improvement when automated resin mixing and mixed solvent system were used. These improvements enable us to produce 48 peptides within a week each of sufficient purity to be used for rapid screening of T‐cell epitopes. ©Munksgaard

ISSN:0367-8377    

DOI:10.1111/j.1399-3011.1996.tb00814.x

出版商:Blackwell Publishing Ltd    

年代:1996

数据来源: WILEY

摘要:

Solid‐phase synthesis of the autoinhibitory domain of calcineurin, CaN A467–491, also produced [aspartimide477]CaN A467–491and [iso‐Asp477]CaNA467–491when Boc‐based chemistry was employed. In addition, the truncated peptide CaN A467‐488was obtained when Fmoc‐based chemistry was employed. All four peptides proved to be effective inhibitors of protein phosphatase activity of calcineurin. The full‐length peptide and theC‐terminally truncated peptide (CaN467‐488) were indistinguishable, withKivalues of 28 ± 3 and 31 ± 5 μM respectively. The internally modified peptides, [iso‐Asp477]CaN A467–491and [aspartimide477]‐CaN A467–491, possessed lower inhibitory potencies (Kivalues of 87 ± 10 and 55 ±3

ISSN:0367-8377    

DOI:10.1111/j.1399-3011.1996.tb00815.x

出版商:Blackwell Publishing Ltd    

年代:1996

数据来源: WILEY

摘要:

A novel ribosome‐inactivating protein (RIP) designated α‐kirilowin was isolated from the seeds ofTrichosanthes kirilowii.The molecular weight of α‐kirilowin was estimated by SDS‐polyacrylamide gel electrophoresis to be 28 800 Da, which is slightly larger than another previously characterized ribosome‐inactivating protein, β‐kirilowin. The amino‐acid composition of α‐kirilowin grossly resembled β‐kirilowin and other ribosome‐inactivating proteins isolated fromT. kirilowiitissues, including trichokirin, trichosanthin and karasurin. Intense immunological cross‐reactivity between the two kirilowins was detected by immunodiffusion. TheN‐terminal sequence of α‐kirilowin was identical to that of β‐kirilowin, at least in the first ten residues. Peptide fingerprinting indicated both kirilowins were closely related. Biological activities as determined by inhibition of protein synthesis in a cell‐free system, suppression of [3H]‐thymidine incorporation into mouse melanoma cells and induction of abortion in mice were very similar for both kirilowins. We propose that the size difference between α‐and β‐kirilowin is either due to aC‐terminal extension in α‐kirilowin or differences in glycosyl

ISSN:0367-8377    

DOI:10.1111/j.1399-3011.1996.tb00816.x

出版商:Blackwell Publishing Ltd    

年代:1996

数据来源: WILEY

摘要:

(φ, ψ) data from crystal structures of 221 proteins having high resolution and sequence similarity cut‐off at the 25% level were analysed by dividing the Ramachandran plot in three regions representing three conformational states: (i) conformational state 1: conformations in the (φ. ψ) range from (‐140°, ‐100°) to (0°, 0°); (ii) conformational state 2: conformations with (φ, ψ) from (‐180°, 80°) to (0°, 180°); and (iii) conformational state 3: all the remaining conformations in the (φ, ψ) plane which are not included in the above two conformational states.Normalized probability values of the occurrence of single amino acid residues in conformational regions 1‐3 and similar values for dipeptides were calculated. Comparisons of single residue and dipeptide normalized probability values have shown that short‐range interactions, although strong, destabilize conformational states of only 44 dipeptides out of the 400 × 9 possible states. However, dipeptide frequency values provide better resolving power than single‐residue potentials when used to predict conformational states of residues in a protein from its primary structure. The simple approach used in the present study to predict conformational states yields an accuracy of>70%, for 14 proteins and an accuracy in the range of 50–700% for 247 proteins. Thus these studies point out yet another use of the Ramachandran plot and the role of tertiary interactions in p

ISSN:0367-8377    

DOI:10.1111/j.1399-3011.1996.tb00817.x

出版商:Blackwell Publishing Ltd    

年代:1996

数据来源: WILEY

摘要:

The solution structure of a synthetic peptide corresponding to residues 151–172 of HIV‐1 integrase has been determined by NMR and CD spectroscopy. Residues 151–172 of HIV‐1 integrase were predicted to be an α‐helix and to be responsible for the oligomerization of HIV‐1 integrase. Two‐dimensional1H NMR and CD studies indicate that this synthetic peptide adopts an amphipathic α‐helical conformation in TFE‐containing solution. However, concentration‐dependent CD studies reveal that this peptide motif does not form dimers or oligomers in solution as predicted. These results are in agreement with the crystal structure of the catalytic domain of HIV‐1 integrase reported rec

ISSN:0367-8377    

DOI:10.1111/j.1399-3011.1996.tb00818.x

出版商:Blackwell Publishing Ltd    

年代:1996

数据来源: WILEY

摘要:

Mass spectrometry was used to determine the molecular mass of rat pituitary β‐endorphin1–31(BErat, 1–31The measured molecular mass (3435 ± 1 Da,n= 5) of endogenous BErat, 1–31differed from the molecular mass of commercially available synthetic BErat, 1–31(3465 ± 1 Da,n= 9), but corresponded to the molecular mass of synthetic BEbovine, 1–31(3436 ± 3 Da,n= 3). Based on the combination of these ESIMS molecular mass measurements, HPLC retention time data, LSIMS measurement of the molecular mass of selected tryptic fragments, and consideration of codon sequences, we suggest that the amino‐acid sequence of endogenous BErat, 1–31differs from the DNA‐deduced sequence of BErat, 1–31, and that endogenous BErat, 1–31contains Ala instead of Val in posi

ISSN:0367-8377    

DOI:10.1111/j.1399-3011.1996.tb00819.x

出版商:Blackwell Publishing Ltd    

年代:1996

数据来源: WILEY